Rat anti pdgfrα

Immunopanning experiment was carried out as previously described with some modifications (Foo, 2013 (link)). The rat or mouse brain was minced and digested with 1 mg/mL papain at 37°C for 1 h. After the cessation of digestion by ovomucoid (Worthington, LS003086), the resulting cell suspension was incubated with primary antibodies [astrocyte, mouse anti-ITGB5 (Invitrogen, 14–0497-82); OPC, rat anti-PDGFRα (BD Biosciences, 558774); OL, mouse anti-PLP (Millipore, MAB388); mouse microglia, rat anti-CD45 (BD, 550539); neuron, rabbit anti-p75 (Abcam, EP1039Y)] diluted in the panning buffer for 30 min at room temperature. The cell suspension was then transferred to a series of secondary antibodies coated 10-cm dishes, and incubated for 30 min. After attachment of the target cell types to the dish, the remnant cell suspension was removed, and the dish was washed with the panning buffer thrice. The cells were then collected with RIPA buffer and subjected to proteomic analysis.

Cell culture on coverslip were fixed with 4% PFA for 8min
and quenched with 10mM Glycine for 30min and then blocked in blocking buffer
(0.02% Triton X-100 and 5% normal donkey serum in PBS) for
20min and incubate with primary antibodies for 40min at room temperature.
Cryosections (12-μm thick) or vibratome sections (50-μm
thick) were permeabilized and blocked in blocking buffer (0.3%
Triton X-100 and 5% normal donkey serum in PBS) for 1 h at room
temperature and overlaid with primary antibodies overnight at 4 °C.
Antibodies used in the study were: rabbit anti-Olig2 (Millipore, AB9610),
rat anti-PDGFRα (BD Bioscience, 558774), mouse anti-APC (CC1,
Oncogene Research, OP80), goat anti-MBP (Santa Cruz Biotechnology,
sc-13914), rabbit anti-CHD8 (Abcam, ab84527), rabbit anti-GFAP (Invitrogen,
#13-0300), rabbit anti-cleaved caspase 3 (Cell Signaling Technology,
#9661), rabbit anti-GFP (Molecular Probes, #A-11122), Rabbit
Monoclonal anti-Sox10 (Abcam, ab180862), mouse anti-NeuN (Millipore,
MAB377). After washing with 0.2% Triton X-100 in PBS, cells or brain
sections were incubated with secondary antibodies conjugated to Cy2, Cy3 or
Cy5 (Jackson ImmunoResearch Laboratories) for 2 h at room temperature,
stained in DAPI for 5 min, washed in PBS and mounted with Fluoromount-G
(SouthernBiotech). Cell counting was carried out in a double-blind
manner.